Raman spectroscopy to assess the differentiation of bone marrow mesenchymal stem cells into a glial phenotype.

Background: Mesenchymal stem cells (MSCs) are multipotent precursor cells with the ability to self-renew and differentiate into multiple cell linage, including the Schwann-like fate that promotes regeneration after lesion. Raman spectroscopy provides a precise characterization of the osteogenic, adipogenic, hepatogenic and myogenic differentiation of MSCs. However, the differentiation of bone marrow mesenchymal stem cells (BMSCs) towards a glial phenotype (Schwann-like cells) has not been characterized before using Raman spectroscopy. Method: We evaluated three conditions: 1) cell culture from rat bone marrow undifferentiated (uBMSCs), and two conditions of differentiation; 2) cells exposed to olfactory ensheathing cells-conditioned medium (dBMSCs) and 3) cells obtained from olfactory bulb (OECs). uBMSCs phenotyping was confirmed by morphology, immunocytochemistry and flow cytometry using antibodies of cell surface: CD90 and CD73. Glial phenotype of dBMSCs and OECs were verified by morphology and immunocytochemistry using markers of Schwann-like cells and OECs such as GFAP, p75 NTR and O4. Then, the Principal Component Analysis (PCA) of Raman spectroscopy was performed to discriminate components from the high wavenumber region between undifferentiated and glial-differentiated cells. Raman bands at the fingerprint region also were used to analyze the differentiation between conditions. Results: Differences between Raman spectra from uBMSC and glial phenotype groups were noted at multiple Raman shift values. A significant decrease in the concentration of all major cellular components, including nucleic acids, proteins, and lipids were found in the glial phenotype groups. PCA analysis confirmed that the highest spectral variations between groups came from the high wavenumber region observed in undifferentiated cells and contributed with the discrimination between glial phenotype groups. Conclusion: These findings support the use of Raman spectroscopy for the characterization of uBMSCs and its differentiation in the glial phenotype.

Afrontamiento a la muerte en estudiantes de medicina del Paraguay en agosto 2021: estudio multicéntrico / Coping with death in Paraguayan medical students in August 2021: a multicenter study

RESUMEN Introducción: el afrontamiento a la muerte constituye una de las situaciones más difíciles y estresantes a las que se tienen que enfrentar los estudiantes de medicina por el contacto cercano a pacientes en riesgo vital. Objetivos: determinar el nivel de afrontamiento a la muerte en estudiantes de medicina. Metodología: se aplicó diseño con enfoque cuantitativo, de tipo observacional, transversal, multicéntrico. Se incluyó a estudiantes de medicina del Paraguay, de ambos sexos, en agosto 2021, que aceptaban participar del estudio. Se utilizó un cuestionario telemático. Se midieron variables sociodemográficas. El nivel de afrontamiento se determinó con la escala de Bugen. La investigación fue aprobada por Comité de Ética de la Universidad Privada del Este, Asunción. Resultados: se incluyeron 504 estudiantes, siendo del sexo femenino en 74 % y con edad media 22 ± 4 años. El 88 % de los encuestados eran de nacionalidad paraguaya (88 %) y 73 % de religión católica. La mayoría nació en Paraguay (88 %), era católica (73 %), se hallaba inscripta como donante de órganos (60 %) y tenía antecedente reciente de muerte de un familiar o amigo (76 %). La escala de Bugen detectó bajo nivel de afrontamiento a la muerte en 50 %, siendo 52 % en las mujeres y 42 % en los varones (p <0,05). Conclusión: el afrontamiento a la muerte en estudiantes de medicina fue bajo. Se sugiere incluir cursos, talleres y planes formativos dentro de las asignaturas o como actividades de extensión para mejorar ese aspecto en el currículo de las facultades de medicina del Paraguay.

ABSTRACT Introduction: coping with death is one of the most difficult tasks that medical students have to face. Objectives: to determine the level of coping with death in medical students. Methodology: we conducted an observational, cross-sectional, multicenter study. We included men and women, medical students from public and private universities from Paraguay in 2021, who agreed to participate in the study. A telematic questionnaire was used. Sociodemographic variables were measured. The level of coping was determined with the Bugen scale. The research was approved by the Comité de Ética of the Universidad Privada del Este, Asunción. Results: 504 students were included, 74 % being female and with a mean age of 22 ± 4 years. Most were born in Paraguay (88 %), were Catholic (73 %), registered as an organ donor (60 %), and had a recent history of the death of a family member or friend (76 %). The Bugen scale detected a low level of coping with death in 52 % of the women and 42 % of the men (p <0.05). Conclusion: coping with death in medical students was low. It is suggested to include courses or workshops to improve this aspect in the curriculum of the medical schools of Paraguay.

Intervención educativa en pacientes con estimación de riesgo de Diabetes mellitus tipo 2 / Educational intervention in patients with estimation of risk of Diabetes mellitus type 2

El reconocimiento de la diabetes de tipo 2 (DM2) como una enfermedad crónica que afecta a millones de personas en el mundo, ha motivado la búsqueda de otros ámbitos de atención de salud, así como enfoques y metodologías que favorezcan un acercamiento real al problema. El objetivo del estudio fue evaluar una intervención educativa en pacientes clasificados de acuerdo a la estimación de riesgo de DM2, Municipio San Diego, Estado Carabobo. Se trató de un estudio descriptivo, de corte longitudinal, de intervención educativa antes-después, con seguimiento de 12 meses y de muestra intencional, que incluyó 80 participantes. Al comparar el nivel de conocimientos y el riesgo de DM2 antes y después de la intervención educativa, se obtuvo diferencias significativas. Se concluye que se produjo un cambio en el nivel de conocimiento de la población estudiada, así como un aumento de la proporción de pacientes que pasaron de un riesgo moderado, a un riesgo bajo de DM2. Los hallazgos encontrados valoran como positiva la intervención educativa y suponen un importante aporte, que evidencia la importancia de la educación en la población.

The recognition of type 2 diabetes (DM2) as a chronic disease affecting millions of people around the world has been a motivation to search for other areas of health care, as well as approaches and methodologies leading to a better management of the problem. The objective of this study was to evaluate an educational intervention in patients classified according to the DM2 risk estimate, from the San Diego Municipality in Carabobo State. This was a descriptive, longitudinal study, of before-after educational intervention, with a 12-month follow-up and an intentional sample, which included 80 participants. When comparing the level of knowledge and risk of DM2 before and after the educational intervention, significant differences were obtained. We conclude that there was a change at the level of knowledge of the studied population, as well as an increase in the proportion of patients who moved from a moderate risk to a low risk of DM2. Findings reveal that educational intervention is positive, as well as an important contribution, which stresses the importance of education in the population.

Characterization of Recombinant Monoclonal Antibody Charge Variants Using OFFGEL Fractionation, Weak Anion Exchange Chromatography, and Mass Spectrometry.

Recombinant monoclonal antibody charge heterogeneity has been commonly observed as multiple bands or peaks when analyzed by charge-based analytical methods such as isoelectric focusing electrophoresis and cation or anion exchange chromatography. Those charge variants have been separated by some of the above-mentioned methods and used for detailed characterization. The utility of a combination of OFFGEL fractionation and weak anion exchange chromatography to separate the charge variants of a recombinant monoclonal antibody was demonstrated in the current study. Charge variants were separated into various fractions of high purity and then analyzed thoroughly by liquid chromatography mass spectrometry. Analysis of intact molecular weights identified the presence of heavy chain leader sequence, C-terminal lysine, and C-terminal amidation. The identified modifications were further localized into different regions of the antibody from analysis of antibody fragments obtained from FabRICATOR digestion. Analysis of tryptic peptides from various fractions further confirmed the previously identified modifications in the basic variants. Asparagine deamidation and aspartate isomerization were identified in acidic fractions from analysis of tryptic peptides. Basic variants have been fully accounted for by the identified modifications. However, only a portion of the acidic variants can be explained by deamidation and isomerization, suggesting that additional modifications are yet to be identified or acidic variants are an ensemble of molecules with different structures.

Detection and quantitation of low abundance oligosaccharides in recombinant monoclonal antibodies.

Oligosaccharides are critical for structural integrity, stability, and biological functions of recombinant monoclonal antibodies. It is relatively easy to characterize, quantify, and determine the impact of major glycoforms. While challenging to detect and quantify, certain low abundance oligosaccharides are highly relevant to the stability and functions of recombinant monoclonal antibodies. Methods were established in this study based on enzymatic digestion to consolidate peaks of the same type of oligosaccharides by removing heterogeneity and thus increase detectability of low abundance peaks. Endo H was used to collapse high mannose oligosaccharides to a single peak of GlcNAc for ease of detection and quantitation. ß-Galactosidase and ß-N-acetylhexosaminidase were used to convert complex oligosaccharides into two peaks containing either GlcNAc2Man3Fuc or GlcNAc2Man3, which simplified the chromatograms and data analysis. More importantly, low abundance hybrid oligosaccharides can only be detected and qualified after ß-galactosidase and ß-N-acetylhexosaminidase digestion. Detection and quantitation of low abundance oligosaccharides can also be achieved using a combination of all three enzymes. These methods can be applied to the development of recombinant monoclonal antibody therapeutics.

Effect of oral hygiene and 0.12% chlorhexidine gluconate oral rinse in preventing ventilator-associated pneumonia after cardiovascular surgery.

BACKGROUND: Ventilator-associated pneumonia (VAP) is a nosocomial infection of multifactorial etiology and has a negative influence on cardiovascular surgery (CVS) outcomes. OBJECTIVES: Determine the effect of toothbrushing plus 0.12% chlorhexidine gluconate oral rinse in preventing VAP after CVS. METHODS: In a quasi-experimental study, patients undergoing heart surgery were enrolled in a protocol for controlling dental biofilm by proper oral hygiene (toothbrushing) and oral rinses with 0.12% chlorhexidine gluconate (Group 1), and they were compared with a historical control group (Group 2), which included patients who underwent cardiac surgery between 2009 and 2010 and who received regular oral hygiene care. Seventy-two hours before surgery, a dentist provided instruction and supervised oral hygiene with toothbrushing and chlorhexidine oral rinses to patients in Group 1. RESULTS: Each group comprised 150 patients. A lower incidence of VAP (2.7% [95% CI 0.7-7.8] vs 8.7% [95% CI 4.9-14.7], P = .04) and a shorter hospital stay (9 ± 3 d [95% CI 8.5-9.5] vs 10 ± 4 d [95% CI 9.4-10.7], P = .01) were observed in Group 1. No significant differences in all-cause in-hospital death were observed between groups (5.3% vs 4.7%, P > .99). The risk for developing pneumonia after surgery was 3-fold higher in Group 2 (3.9, 95% CI 1.1-14.2). CONCLUSIONS: Oral hygiene and mouth rinses with chlorhexidine under supervision of a dentist proved effective in reducing the incidence of VAP.

Association between oral health and acute coronary syndrome in elderly people.

UNLABELLED: With the aim of contributing information to help clarify the association between oral health, periodontal disease and ischemic cardiomyopathy, a comparative study was performed on hospitalized patients with and without acute coronary syndrome (ACS), evaluating atherogenic risk factors (ARF), level of oral hygiene and dental and periodontal health status. The study included patients in the coronary unit with ACS and patients in regular floor bed without evidence of cardiovascular pathology at Hospital Español, Buenos Aires, Argentina. The following ARFs were analyzed for all patients: hypertension, cholesterol, diabetes, obesity and smoking. The clinical dental examination included recording dental charts and variables related to oral hygiene, epidemiological indices and diagnosis of periodontal disease. The data collected were used to compare the ACS Group to the Control Group regarding prevalence of the ARFs and clinical dental variables studied. A total of 146 patients were studied: 81 male and 65 female, with average age 69.8 +/- 10.14 years. Prevalence of hypercholesterolemia was found to be significantly higher in patients in the ACS group than in the control group (p = 0.043). The other ARFs considered did not differ. The levels of oral hygiene were similar in both study groups. The prevalence of decayed and filled teeth was similar in both groups, although there was a significantly higher number of missing teeth in the ACS group (p < 0.001). Patients with ACS had more severe periodontal disease and higher levels of gingival inflammation than the control group (p < 0.001). CONCLUSION: Oral health of patients with ACS was worse than oral health of patients without cardiovascular disease. The difference was shown by greater severity of periodontal disease in patients with ACS even though the level of oral hygiene was similar in both groups.

Association between oral health and acute coronary syndrome in elderly people

Con el objetivo de aportar información que permita esclarecer la relación entre la salud bucal, la enfermedad periodontal y la cardiopatía isquémica, se efectuó un estudio comparativoen pacientes internados con y sin síndrome coronario agudo (SCA) evaluando factores de riesgo aterogénicos (FRA), nivel de higiene bucal, estado de salud dental y periodontal. Se incorporaron al estudio pacientes internados en unidad coronaria con SCA y pacientes internados en sala general sin evidencia de patología cardiovascular del Hospital Español, Buenos Aires, Argentina. En todos los pacientes se consignaronlos FRA: hipertensión arterial, colesterol, diabetes, obesidad ytabaquismo. En el examen clínico odontológico se registró: odontograma, variables relacionadas con la higiene bucal, se confeccionaron índices epidemiológicos y se realizó el diagnóstico del cuadro periodontal. Con los datos recolectados, se realizó un examen comparativo entre los dos grupos de estudio (Grupo con SCA vs Grupo Control) respecto a la prevalenciade los FRA y las variables clínicas odontológicas estudiadas. Se estudiaron 146 pacientes, 81 hombres - 65 mujeres, con promedio de edad de 69.8±10.14 años. Se observó que laprevalencia de hipercolesterolemia fue significativamente mayor en los pacientes del grupo SCA respecto a los controles (p=0.043). Los restantes FRA considerados no mostraron diferencias. Los niveles de higiene bucal fueron semejantes entre ambos grupos de estudio. El grupo SCA tuvo similar prevalencia de caries y restauraciones respecto al grupo Control, sin embargo, perdieron significativamente más piezasdentarias (p<0.001). Los enfermos con SCA tuvieron cuadros periodontales más severos y niveles de inflamación gingivalsuperiores en comparación con el grupo control (p<0.001). Conclusión: El estado de salud bucal de los pacientes con SCA fue peor respecto de aquellos pacientes que no presentabanenfermedad cardiovascular.

Niveles séricos de la proteína transportadora de esteres de colesterol (CETP) y perfil lipídico de pacientes diabéticos tipo 2: unidad de diabetes Ciudad Hospitalaria Dr. Enrique Tejera / Serum levels of cholesterol ester transfer protein (CETP) and lipid profile of type 2 diabetic patients: diabetes unit Hospital City Dr. Enrique Tejera

La Proteína Transportadora de Esteres de Colesterol (CETP), enzima que juega un rol importante en el transporte reverso del colesterol, ha sido asociada con aterosclerosis; sin embargo no se tiene claro su papel en las alterciones del perfil lipídico de pacientes diabéticos tipo 2. Por este motivo en el presente estudio se determinaron los niveles séricos de CETP, y el perfil lipídico en 33 pacientes de sexo masculino con diabetes tipo 2, en edades coprendidas entre 40 y 65 años, que asistieron a la consulta de la Unidad de diabetes de la CHET. El grupo control estuvo constituidos por 30 sujetos no diabéticos, normolipémicos, del mismo sexo y en el mismo rango de edad. La concentración sérica de CETP fue determinada por inmunoensayo de fase sólida (ELISA) utilizando un kit comercial, el cual no reporta valores de referencia por ser sólo para uso de investigación; y el perfil lipídico por métodos enzimáticos-colorimétricos. Para el grupo de diabéticos la concentración promedio ± desviación estándar de CETP fue 2,4 ± 0,56 µ/mL y para los controles 2,63 ± 0,7 µ/mL, encontrándose ambos dentro de los valores reportados por otros investigadores no observándose diferencia significativa entre ambos grupo <0,05. En cuanto al perfil lipídico los pacientes diabéticos mostraron valores normales, con promedios de 183,9 ± 32,45 mg/dL para el colesterol, 44,46 ± 7,05 mg/dL para HDL-c, 114,61 ± 10,50 mg/dL para, LDL-c y 382 ± 3,93 mg/dL para las VLDL-c, observándose sólo un aumento en los niveles de triglicéridos con un promedio de 211,48 ± 137,54 mg/dL (X ± DE). Para el grupo control dichos niveles se encontraron dentro del rango de referencia. La alteración del perfil lipídico observada en los pacientes diabéticos tipo II (aumento de los triglicéridos) no guardó relación con los niveles séricos de CETP